|Entry||Database: EMDB / ID: EMD-4086|
|Title||RNA polymerase I-Rrn3 complex|
|Sample||S. cerevisiae RNA polymerase I in complex with activating factor Rrn3|
|Biological species||Saccharomyces cerevisiae (baker's yeast)|
|Method||single particle reconstruction / cryo EM / Resolution: 7.7 Å|
|Authors||Torreira E / Louro JA / Gil-Carton D / Gallego O / Calvo O / Fernandez-Tornero C|
|Citation||Journal: Elife / Year: 2017|
Title: The dynamic assembly of distinct RNA polymerase I complexes modulates rDNA transcription.
Authors: Eva Torreira / Jaime Alegrio Louro / Irene Pazos / Noelia González-Polo / David Gil-Carton / Ana Garcia Duran / Sébastien Tosi / Oriol Gallego / Olga Calvo / Carlos Fernández-Tornero /
Abstract: Cell growth requires synthesis of ribosomal RNA by RNA polymerase I (Pol I). Binding of initiation factor Rrn3 activates Pol I, fostering recruitment to ribosomal DNA promoters. This fundamental ...Cell growth requires synthesis of ribosomal RNA by RNA polymerase I (Pol I). Binding of initiation factor Rrn3 activates Pol I, fostering recruitment to ribosomal DNA promoters. This fundamental process must be precisely regulated to satisfy cell needs at any time. We present in vivo evidence that, when growth is arrested by nutrient deprivation, cells induce rapid clearance of Pol I-Rrn3 complexes, followed by the assembly of inactive Pol I homodimers. This dual repressive mechanism reverts upon nutrient addition, thus restoring cell growth. Moreover, Pol I dimers also form after inhibition of either ribosome biogenesis or protein synthesis. Our mutational analysis, based on the electron cryomicroscopy structures of monomeric Pol I alone and in complex with Rrn3, underscores the central role of subunits A43 and A14 in the regulation of differential Pol I complexes assembly and subsequent promoter association.
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_4086.map.gz / Format: CCP4 / Size: 16.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.77 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire S. cerevisiae RNA polymerase I in complex with activating factor Rrn3
|Entire||Name: S. cerevisiae RNA polymerase I in complex with activating factor Rrn3|
Number of components: 1
-Component #1: protein, S. cerevisiae RNA polymerase I in complex with activatin...
|Protein||Name: S. cerevisiae RNA polymerase I in complex with activating factor Rrn3|
Recombinant expression: No
|Mass||Theoretical: 594 kDa|
|Source||Species: Saccharomyces cerevisiae (baker's yeast)|
|Specimen||Specimen state: Particle / Method: cryo EM|
|Sample solution||Specimen conc.: 0.06 mg/mL / pH: 7.8|
|Vitrification||Instrument: FEI VITROBOT MARK III / Cryogen name: ETHANE / Temperature: 277 K / Humidity: 95 %|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 68 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Magnification: 47000.0 X (nominal), 79096.0 X (calibrated) / Cs: 2 mm / Imaging mode: BRIGHT FIELD / Defocus: 1900.0 - 4200.0 nm|
|Specimen Holder||Model: FEI TITAN KRIOS AUTOGRID HOLDER|
|Camera||Detector: FEI FALCON II (4k x 4k)|
|Image acquisition||Number of digital images: 1288|
|Processing||Method: single particle reconstruction / Number of projections: 32175|
|3D reconstruction||Software: RELION / Resolution: 7.7 Å / Resolution method: FSC 0.143 CUT-OFF|
|FSC plot (resolution estimation)|
-Atomic model buiding
|Modeling #1||Refinement protocol: rigid body / Refinement space: REAL / Overall bvalue: 537|
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